A) Amino acid alignment of KL1 and KL2 domains of sKlotho. Arg148 of KL1 is conserved in KL2 domain (green box). His246, Ser299, and motif 465GluTrpHisArg468 of KL1 domain are not conserved in KL2 (red box). B) Structural differences between KL1 and KL2 domain of sKlotho for residues implicated in binding to α2-3-sialyllactose (Arg148, His246, and motif 465GluTrpHisArg468). KL1 residues are green. Closest matching residues from KL2 domain (turquoise) for same KL1 residues show considerable local differences in structure, giving plausible reason why KL2 domain may be unable to bind α2-3-sialyllactose.